Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.5.1.52 (PNGase F)
 1,527 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Spodoptera frugiperda cells are the standard host system for baculovirus-vector-mediated expression of recombinant glycoproteins. In an attempt to explore their ability to produce complex N-glycans containing terminal neuraminic acid, we tested both mock- and AcMNPV-infected SF9 cells. To elucidate the structures of the carbohydrate chains of cellular glycoproteins, radiolabeled oligosaccharides were liberated by treatment with endo-H and glycopeptidase F. When the endo-H resistant material was subjected to sequential degradation with exoglycosidases, only truncated carbohydrates with the structures Man3GlcNAc2 and Man3[Fuc]GlcNAc2 were found. There was no evidence for the presence of neuraminic acid and complex N-glycans. The results indicate that SF9 cells have only a limited capacity to process N-glycans. Infection with AcMNPV has no significant effect on glycosylation in these cells.
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PMID:Baculovirus infection does not alter N-glycosylation in Spodoptera frugiperda cells. 807 3

For overexpression of the N-methyl-D-aspartate (NMDA) receptor subunit 1b (NMDAR1b), its corresponding cDNA was extended by codons for six histidine residues at the 3'-end, cloned into a baculovirus transfer vector and integrated into the viral genome. Infection of Trichoplusia ni insect cells (High FiveTM cells) with recombinant baculovirus resulted in the production of 126- and 105-kDa NR 1b proteins in the cell membrane fraction. Enzymatic deglycosylation with PNGase F as well as infection of the insect cells in the presence of tunicamycin revealed that the two proteins represented the N-glycosylated and non-glycosylated forms of NMDAR1b, respectively. The recombinant NR1b protein was also identified with immunocytochemical methods employing a monoclonal antibody which recognized the six histidine residues. The affinity of this histidine tag to nickel ions was used for the purification of the NR1b protein. The glycine binding site of the subunit was successfully identified and analyzed with the specific antagonist 5,7-[3-3H]dichlorokynurenate (DCKA). The observed binding characteristics were similar to those obtained for native NMDA receptors. Whereas in electrophysiological measurements a functional NMDA receptor channel could not be found in infected insect cells, its expression was demonstrated in the Xenopus oocyte system after injection of the NMDAR1b cDNA construct.
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PMID:Overexpression of a functional NMDA receptor subunit (NMDAR1) in baculovirus-infected Trichoplusia ni insect cells. 888 56